Wellcome Trust Sanger Institute scientists and their collaborators have developed a new analysis tool that was able to show, for the first time, which genes were expressed by individual cells in different genetic versions of a benign blood cancer.

Single cell RNA sequencing can define cell types by revealing differences in the proteins produced by individual cells, however analysing the data remains challenging. Reported in Nature Methods today, the new open source computer tool called Single Cell Consensus Clustering (SC3) was shown to be more accurate and robust than existing methods of analysing single-cell RNA sequence data, and is freely available for researchers to use*.

Recent advances in single-cell genomics technology has made it possible to separate individual cells from different tissues and organs, and measure the sets of RNA messages – called the transcriptome – which help give each cell its own identity. These individual transcriptomes can be used to define cell types and to understand the functions of healthy and diseased cells in the human body. This technology has enormous potential for biological research.

In order to analyse the transcriptomic data, similar cells need to be grouped together. However, it is hard to know what criteria to use to group them, and the data is often very complex. The researchers developed the SC3 computer tool to overcome these problems and validated it using several publicly available gold standard datasets.

Dr Vladimir Yu Kiselev, first author from the Sanger Institute, said "We created the new SC3 tool to analyse complex single-cell RNA-sequence data, and showed that it is more robust and accurate than existing methods at grouping cells. The SC3 tool contains added features that help interpret the biological function of the cells in that group, such as lists of marker genes for each group. We expect this will be used by many researchers around the world."

The SC3 tool was then used to analyse single-cell RNA-sequence data from two patients diagnosed with myeloproliferative neoplasm (MPN) blood cancers. Pre-malignant MPN occurs when the bone marrow makes too many blood cells, and in 10 per cent of patients can lead to overt leukaemia.

Patients often have multiple versions of the cancer, called subclones, which have different mutations, and the researchers wanted to find if the expression levels of RNA correlated with the different mutations. Previous attempts to analyse the RNA datasets with other methods had failed, however SC3 was able to resolve the datasets and showed that each cancer-causing mutation led to different proteins being expressed.

Prof Tony Green, an author from the Wellcome Trust-MRC Stem Cell Institute and Cambridge University, said: "The SC3 tool was able to use patterns of gene expression to distinguish, within an individual cancer, subclones that carried different mutations. This approach will help us define the cellular heterogeneity within each cancer, an important step towards improving cancer treatment".

Dr Martin Hemberg, lead author on the paper from the Wellcome Trust Sanger Institute, said: "It has been difficult to fully exploit single-cell RNA-sequence data due to the current lack of computational methods for analysing them. Our study shows that SC3 is an accurate and user-friendly tool, which can analyse complex datasets. We hope that this tool will help researchers gain new biological insights from transcriptome datasets in the future and provide information for diseases that affect specific cell types."

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Notes to editors:

*SC3 is available from http://www.bioconductor.org/packages/SC3/ and the source code can be found at https://github.com/hemberg-lab/SC3

Selected Websites:

The Wellcome Trust-Medical Research Council Stem Cell Institute

The Wellcome Trust-Medical Research Council Stem Cell Institute explores and defines the properties of stem cells to establish their true medical potential. Leading research scientists, technology specialists and doctors work side by side to create a world-leading centre of excellence in stem cell biology and medicine. The Institute also provides high level training for young researchers from around the world and collaborates with bio-industry. http://www.stemcells.cam.ac.uk/

The University of Cambridge

The mission of the University of Cambridge is to contribute to society through the pursuit of education, learning and research at the highest international levels of excellence. To date, 96 affiliates of the University have won the Nobel Prize. Founded in 1209, the University comprises 31 autonomous Colleges, which admit undergraduates and provide small-group tuition, and 150 departments, faculties and institutions. Cambridge is a global university. Its 19,000 student body includes 3,700 international students from 120 countries. Cambridge researchers collaborate with colleagues worldwide, and the University has established larger-scale partnerships in Asia, Africa and America. The University sits at the heart of one of the world's largest technology clusters. The 'Cambridge Phenomenon' has created 1,500 hi-tech companies, 14 of them valued at over US$1 billion and two at over US$10 billion. Cambridge promotes the interface between academia and business, and has a global reputation for innovation. http://www.cam.ac.uk

The Wellcome Trust Sanger Institute

The Wellcome Trust Sanger Institute is one of the world's leading genome centres. Through its ability to conduct research at scale, it is able to engage in bold and long-term exploratory projects that are designed to influence and empower medical science globally. Institute research findings, generated through its own research programmes and through its leading role in international consortia, are being used to develop new diagnostics and treatments for human disease. http://www.sanger.ac.uk

Wellcome

Wellcome exists to improve health for everyone by helping great ideas to thrive. We're a global charitable foundation, both politically and financially independent. We support scientists and researchers, take on big problems, fuel imaginations and spark debate. http://www.wellcome.ac.uk